Sanitary swab collection system, microfluidic assay device, and methods for diagnostic assays

ABSTRACT

Biohazard specimen collection containers are provided with an external disposable skin, that is stripped away and discarded after the biohazardous specimen is collected, thus reducing or eliminating objectionable or dangerous residues on the outside surfaces of the container. Further, we teach that the sample collection container with external disposable skin may also serve as an integrated microfluidic biosample processing and analytical device, thereby providing a single entry, disposable assay unit, kit and system for “world-to-result” clinical diagnostic testing. These integrated assay devices are provided with synergic, multiple safe-handling features for protecting healthcare workers who handle them. The modified collection containers and analytical devices find application, for example, in PCR detection of infectious organisms or pathogenic markers collected on a swab.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.12/695,487 filed Jan. 28, 2010, now allowed; which application is acontinuation of International Patent Application No. PCT/US2008/071810filed Jul. 31, 2008; which application claims the benefit under 35U.S.C. §119(e) of U.S. Provisional Application No. 60/953,045, filedJul. 31, 2007; which applications are incorporated herein by referencein their entireties.

STATEMENT OF GOVERNMENT INTEREST

This invention was made with government support under Contract No. U01AI070801 awarded by National Institutes of Health. The government hascertain rights in this invention.

BACKGROUND

1. Technical Field

The invention relates to medical and veterinary sample collectiondevices and to medical and veterinary analytical devices of specializedform and function, and to integrated microfluidic devices for bothsample collection and analysis. The invention further relates to amethod for biohazard sample collection.

2. Description of the Related Art

The art relating to handling of swabs is well established, but remainsin need of improvement, both to ensure the integrity of the clinicalsample and its protection from contamination, but also to ensure thathealthcare professionals are not unnecessarily or inadvertently exposedto biological material on the exterior surfaces of the swab container.Once the external surfaces are contaminated during sample collection,exposure readily occurs when a swab container is passed from hand tohand, and no on-board means is known to refresh or cleanse the outsidesurfaces of the sample container.

We have reviewed the patent literature, and found little or no teachingthat comments on this problem. U.S. Pat. No. 4,803,998 to Kezes relatesto a swab retaining vial cap and describes a combination containmentvial with cap and with swab mounted inside the cap, the vial containinga medium for preserving a sample on the swab during shipment. The swabis removed from the cap to collect a sample and the swab tip can then bebroken off when inserted into the vial so that the swab tip drops to thebottom of the vial without contamination by the user. The cap is thensealed. FIG. 4 shows a swab with frangible shaft. The patent isindicative of early efforts to protect a sample from contamination. Thisseems to accurately reflect the overall state of the art as it exists atthis filing. We note that while the interior of the vial is carefullyprotected from contamination, the exterior is subject to contaminationduring handling, and becomes a fomite vector for infectious disease.Samples collected in this way are frequently removed for analysis at aseparate location, and those who handle the sample container mayinadvertently be exposed to material on the exterior surface of thesample container.

U.S. Pat. No. 6,991,898 to O'Connor (Jan. 31, 2006) describes aself-contained diagnostic test device for collection and detection of ananalyte in a biological specimen. The device comprises a tubular swaband reagent dispensing cap. The reagent dispensing cap delivers one ormore selected reagents to an assay chamber upon the rotation of thereagent chamber.

In U.S. Pat. No. 7,098,040 to Kaylor, a swab-based diagnostic testdevice is provided. The test device contains a reagent and a rupturableseal for adding the reagent to the sample after the swab is sealedinside the device.

U.S. Pat. No. 6,277,646 to Guirguis provides a device for bothcollecting and testing a fluid specimen. A fluid specimen is collectedand an aliquot is transferred to an isolation chamber, from which a flowpath to a test chamber is opened.

U.S. Pat. No. 6,565,808 to Hudak describes a fluid flow actuating deviceor structure, such as a valve, which separates the sample receivingchamber from the test platform. The test method involves collecting asample, contacting the sample with the proprietary test device, anddetecting the analyte in the sample.

U.S. Pat. No. 6,248,294 to Nason relates to a self-contained diagnostictest unit for use in the collection and analysis of a biologicalspecimen. The test unit comprises is tubular housing for capturing aswab. A reagent dispenser cap delivers reagents to the specimen chamberand a diagnostic strip assembly is mounted on the housing so a portionof the specimen can flow by wick action through the test strip,producing a visible color change.

U.S. Pat. Nos. 5,266,266 and 5,879,635 to Nason relate to a reagentdispenser which includes a pair of reagent chambers with selectedreagents therein, and a dual nib for hermetically sealing the reagentchambers. A portion of the dispenser is deformable to break or otherwiseto displace the nib in a manner permitting the two reagents to flowtogether and mix within one of the reagent chambers. The deformableportion or the dispenser can then be squeezed to express the mixedreagents for delivery to contact the specimen to be analyzed. In apreferred form, the dispenser is a cap assembly on an open-ended tubularhousing configured for receiving a swab.

Similarly, U.S. Pat. No. 6,890,484 to Bautista relates to in-line testdevice and describes a swab receiving port integrated into the body of alateral flow strip. No means for protecting the exterior of the testapparatus is described. Goodfield, in Sampling and Assay Device(WO1997/23596), discloses a swab and swab container with liquid assayreagents accessible by rupture of foil liners, again with no outerdisposable protective layer.

All the above devices and methods are deficient for the present purposein that the operator is exposed to contamination of the externalsurfaces of the specimen collection container by contact with residuesof specimen or unrelated patient-derived bodily material, which may beunhygienic and grossly objectionable. This problem is apparently notconsidered.

United States Patent Application 2005/0009200 to Guo relates to asanitary and compact fecal occult blood collector kit. The swab tip inthis case is covered “for hygienic purposes”. Also disclosed is apackage for the swab and the cover. However, on closer study, thepurpose of the cover is again to protect the sample, not the handle ofthe swab contacted by the operator or the external surfaces of the swabcollection container, and the exterior of the package cannot be cleanedof contaminating matter that accumulates during sampling. Further, theswab must again be retrieved from the package. Thus while the sample isprotected, the user is potentially exposed at multiple levels.

Miniaturizing some of the processes involved in clinical analyses,including nucleic acid, immunological and enzymatic analysis, orcombinations thereof, has been achieved using microfluidic devices.Microfluidic techniques known in the art include electrophoreticdetectors, for example those designed by ACLARA BioSciences Inc., or theLabChip™ by Caliper Technologies Inc, and hybridization detectors suchas those manufactured by Nanogen of San Diego. Also indicative of thestate of the art are PCT Publication WO1994/05414, U.S. Pat. Nos.5,498,392, 5,304,487, 5,296,375, 5,856,174, 6,180,372, 5,939,312,5,939,291, 5,863,502, 6,054,277, 6,261,431, 6,440,725, 5,587,128,5,955,029, 5,498,392, 5,639,423, 5,786,182, 6,261,431, 6,126,804,5,958,349, 6,303,343, 6,403,037, 6,429,007, 6,420,143, 6,572,830,6,541,274, 6,544,734, 6,960,437, 6,762,049, 6,509,186, 6,432,695,7,018,830, and 2001/0046701, 2003/0138941, and International Pat. Nos.WO 2003/004162, WO2002/18823, WO2001/041931, WO1998/50147, WO1997/27324,all of which describe apparatuses and methods incorporating variousmicrofluidic processing and analytical operations involved in nucleicacid analysis, and are incorporated herein by reference.

Co-assigned to Micronics, Inc of Redmond Wash., and also incorporatedherein in full by reference, are U.S. Pat. No. 6,743,399 (“PumplessMicrofluidics”), U.S. Pat. No. 6,488,896 (“Microfluidic AnalysisCartridge”), U.S. Pat. No. 5,726,404 (“Valveless Liquid Microswitch”),U.S. Pat. No. 5,932,100 (“Microfabricated Differential Extraction Deviceand Method”), (“Tangential Flow Planar Microfluidic Fluid Filter”), U.S.Pat. No. 5,872,710 (“Microfabricated Diffusion-Based Chemical Sensor”),U.S. Pat. No. 5,971,158 (“Absorption-Enhancing Differential ExtractionDevice”), U.S. Pat. No. 6,007,775 (“Multiple Analyte Diffusion-BasedChemical Sensor”), U.S. Pat. No. 6,581,899 (“Valve for Use inMicrofluidic Structures”), U.S. Pat. No. 6,431,212 (“Valve for Use inMicrofluidic Structures”), U.S. Pat. No. 7,223,371 (“MicrofluidicChannel Network Device”), U.S. Pat. No. 6,541,213 (“Microscale DiffusionImmunoassay”), U.S. Pat. No. 7,226,562 (“Liquid Analysis Cartridge”),U.S. Pat. No. 5,747,349 (“Fluorescent Reporter Beads for FluidAnalysis”), US Patent Applications 2005/0106066 (“Microfluidic Devicesfor Fluid Manipulation and Analysis”), 2002/0160518 (“MicrofluidicSedimentation”), 2003/0124619 (“Microscale Diffusion Immunoassay”),2003/0175990 (“Microfluidic Channel Network Device”), 2005/0013732(“Method and system for Microfluidic Manipulation, Amplification andAnalysis of Fluids”), 2007/0042427, “Microfluidic Laminar Flow DetectionStrip”, 2005/0129582 (System and Method for Heating, Cooling and HeatCycling on a Microfluidic Device); and unpublished US Patent documentstitled, “Integrated Nucleic Acid Assays,” “Microfluidic Cell Capture andMixing Circuit”, “Microfluidic Mixing and Analytical Apparatus,” “Systemand Method for Diagnosis of Infectious Diseases”, “Methods and Devicesfor Microfluidic Point of Care Assays”, “Integrated Microfluidic AssayDevices and Methods”, and “Microscale Diffusion Immunoassay UtilizingMultivalent Reactants”, all of which are hereby incorporated in full byreference. Also representative of microfluidic technologies that areco-assigned to Micronics are PCT Publications WO 2006/076567 and2007/064635, all incorporated herein in full by reference for what theyenable.

The utility and breadth of microfluidic assays for nucleic acid assaysis further demonstrated in the scientific literature, the teachings ofwhich are incorporated by reference herein. These teachings include, forexample, Nakano H et al. 1994. High speed polymerase chain reaction inconstant flow. Biosci Biotechnol Biochem 58:349-52; Wilding, P et al.1994. PCR in a silicon microstructure. Clin Chem 40(9):1815-18; WoolleyA T et al. 1996. Functional integration of PCR amplification andcapillary electrophoresis in a microfabricated DNA analysis device. AnalChem 68:4081-86; Burke D T et al. 1997. Microfabrication technologiesfor integrated nucleic acid analysis. Genome Res 7:189-197; Kopp et al.1998. Chemical amplification: continuous-flow PCR on a chip. Science280:1046-48; Burns, M A. 1998. An Integrated Nanoliter DNA AnalysisDevice. Science 282:484-87; Belgrader P et al. 1999. PCR Detection ofbacteria in seven minutes. Science 284:449-50; Lagally E T et al. 2001.Fully integrated PCR-capillary electrophoresis microsystem for DNAanalysis. Lab Chip 1:102-07; Tudos A J et al. 2001. Trends inminiaturized total analysis systems for point-of-care testing inclinical chemistry. Lab Chip 1:83-95; Belgrader P et al. 2002. Abattery-powered notebook thermocycler for rapid multiplex real-time PCRanalysis. Anal Chem 73:286-89; Hupert L M et al. 2003. Polymer-BasedMicrofluidic Devices for Biomedical Applications. In, (H Becker and PWoias, eds) Microfluidics, BioMEMS, and Medical Microsystems, Proc SPIEVol 4982:52-64; Chartier I et al. 2003. Fabrication of an hybridplastic-silicon microfluidic device for high-throughput genotyping. In,(H Becker and P Woias, eds) Microfluidics, BioMEMS, and MedicalMicrosystems, Proc SPIE Vol 4982:208-219; Anderson R C et al. 2000. Aminiature integrated device for automated multistep genetic assays. NuclAcids Res 28(12):[e60,i-vi]; Yang, J et al. 2002. High sensitivity PCRassay in plastic micro reactors. Lab Chip 2:179-87; Giordano B C et al.2001. Polymerase chain reaction in polymeric microchips: DNAamplification in less than 240 sec. Anal Biochem 291:124-132; KhandurinaJ et al. 2000. Integrated system for rapid PCR-based DNA analysis inmicrofluidic devices. Anal Chem 72:2995-3000; Chiou, J et al. 2001. AClosed-Cycle Capillary Polymerase Chain Reaction Machine. Anal Chem73:2018-21; Yuen, P K et al. 2001. Microchip module for blood samplepreparation and nucleic acid amplification reactions. Genome Res11:405-412; Zhou X, et al. 2004. Determination of SARS-coronavirus by amicrofluidic chip system. Electrophoresis. 25(17):3032-9; Liu Y et al.2002. DNA amplification and hybridization assays in integrated plasticmonolithic devices. Anal Chem 74(13):3063-70; Zou, Q et al. 2002.Micro-assembled multi-chamber thermal cycler for low-cost reaction chipthermal multiplexing. Sensors Actuators A 102:224-121; Zhang C et al.2006. PCR Microfluidic devices for DNA amplification. Biotech Adv24:243-84, and Zhang, C and Xing D. 2007. Miniaturized PCR chips fornucleic acid amplification and analysis: latest advances and futuretrends. Nucl Acids Res 35(13):4223-37.

Thus there is a clear and ongoing interest in microfluidic devices forclinical and veterinary diagnostic assays. As these commercialapplications increase, the world-to-chip interface is receivingincreasing attention, and we note that little has been done in the areaof sample collection to both improve the validity of nucleic acidamplifications by preventing cross-sample contamination, and just asimportantly, to prevent exposure of those persons handling the specimensto objectionable or potentially infectious materials. As has been noted,(Nelson, D. B. et al. 2003. “Self-Collected Versus Provider-CollectedVaginal Swabs for the Diagnosis of Bacterial Vaginosis: an Assessment ofValidity and Reliability,” J Clin Epidemiol, 56:862-866), there is anincreasing trend toward patient self-collection of samples, often withswabs or cups. Typically the patient is not provided with means toensure that the external surfaces of the sample collection device doesnot become contaminated with the sample or related biological fluidsduring handling. These swabs or cups are typically then processed orhandled by ungloved couriers and paraprofessionals and must then betransferred to the analytical device or further handled and processed bynursing and laboratory personnel. The sample collection device thusbecomes a fomite potentially capable of spreading infectious disease tonumerous persons, and a method or means for eliminating or at leastreducing the exposure of health workers to the contaminated exterior ofthe sample collection vials, bottles, cups, tubes, and so forth, hasbeen a longstanding and unmet need in the healthcare industry.

Furthermore, awareness of the dangers of unsafe handing of biologicalfluids and specimens has increased dramatically in the last two decades,and single-entry devices are increasingly needed that seamlesslyintegrate sample preparation, extraction, and analysis withoutunnecessary operator exposure. A further objective we have identified isthe need to fully integrate the device into a disposable format, so thatonce a sample is collected, either by patient or by a healthprofessional, all remaining steps of the analysis, up to and includingdisplay of the result, are performed without further personal exposureto the sample. A critical step in this process is thus the refreshing ordisinfecting of the external surface sample collection container(whether it is also the analytical device or not), and to our knowledge,satisfactory solutions to this problem have not been recognized orbrought forward prior to our disclosure herein.

BRIEF SUMMARY

Swabs are extremely useful for collecting specimens. Followingcollection of a specimen on a swab, the swab must be generally protectedduring subsequent transport and processing for analysis. During initialhandling, contamination of the external surfaces of the swab collectioncontainer by contact with residues of specimen or unrelatedpatient-derived bodily material, which may be unhygienic and grosslyobjectionable, is almost inevitable. Gloves are protective only to thehands on which they are worn, not to the swab collection container. Wesee a solution to this problem as an unrecognized and unmet need withsignificant potential benefits, particularly in reduction of nosocomialinfections, for example, and more generally in reduction of diseasetransmission to health care workers, and also in improvement of samplequality, which is mandatory for tests such as PCR, where false positivesdue to cross-contamination will invalidate any testing system.

Cross-contamination by transmission on the surfaces of fomites is alongstanding problem. We find that this problem can be alleviated orsignificantly reduced by applying a disposable external skin to thecollection device, and by removing the skin after the risk of exposureto further contamination is ended. Contamination risk is most greatduring the act of specimen collection itself, and decreases greatlyafter the specimen collection container is removed from the samplingsite. Contamination of the external surfaces of an article passed fromhand-to-hand, or hand-to-machine, with normal flora and normal squamousepithelial cells, is significantly less likely to result in falsediagnostic positives for a pathogenic condition.

We disclose a biohazard swab collection device or container, comprisinga body with external surfaces, an internal hollow volume, and a sealableclosure for separating said external surfaces from said internal hollowvolume, said external surface further comprising a disposable externalskin layer, whereby after the biohazardous swab is enclosed and sealedwithin the internal hollow volume, any biohazardous residues accumulatedon the external surfaces are removed by removing and disposing of thedisposable external skin layer, and further optionally comprising avalve separating said internal hollow volume into a swab receivingchamber and a microfluidic assay circuit with a microfluidic channel andan on-board liquid reagent.

We further disclose a method wherein the specimen is not limited to aswab and the specimen collection device is not limited to an analyticaldevice. The general method comprises the steps of:

a) providing a sample and a specimen collection container with body andwith sample receiving orifice, said body with external surface andinternal hollow volume, said external surface with disposable skin orskins, said sample receiving orifice with sealable closure;

b) inserting said sample into said sample receiving orifice;

c) closing said sealable closure said sample receiving orifice; therebycapturing said sample; and,

d) removing said disposable skin or skins from said external surface;thereby renewing said external surface.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a perspective view of a representative specimen collectiondevice with external skins and with integrated sample processing andanalytical assay capability.

FIG. 2 is a perspective view of a sample swab with frangible handle.

FIG. 3A is a plan view of the upper surface of the device of FIG. 1, andshows section plane 3B. FIG. 3B is a section of the device of FIG. 1 onplane 3B, and shows the swab receiving chamber and inner workings of anembodiment of the integrated device. Representative inner workings areindicated schematically.

FIG. 4 is an exploded view of protective external disposable skinsapplied to a representative specimen collection device.

FIG. 5 is a conceptual illustration of the manufacture of a heat-shrinkexternal disposable skin on a representative specimen collection device.

FIG. 6 is an illustration of the assembly of a disposable bag applied toa representative specimen collection device.

FIG. 7 is an exploded view of a Styrofoam or composite coverblockassembly applied to a representative specimen collection device.

FIG. 8 is a sketch of a device with composite cover formed in place overand around the device.

FIGS. 9A-E is a sequential view of the steps of a method in which arepresentative specimen collection device fitted with a disposableexternal sanitary skin is used to collect a specimen on a swab.

FIG. 10 is a block diagram of the steps of a method for collecting abiohazardous swab in a swab collection device fitted with a disposableexternal sanitary skin.

FIG. 11 is a block diagram of the more general steps of a method forcollecting a biohazardous specimen in a specimen collection devicefitted with a disposable external sanitary skin.

FIGS. 12A and B show a detail of a tab on the disposable external coverfor use in removing the protective cover after the specimen iscollected.

FIG. 13 is a second embodiment of a specimen collection device for aswab or tampon, and includes operator interface and real-timepoint-of-care data display that is hidden under the cap of a protectiveremovable overlayer during specimen collection.

FIG. 14 is a section down the long axis of a third embodiment of aspecimen collection device for a swab or tampon, and includes operatorinterface and real-time point-of-care data display that is hidden undera protective removable overlayer during specimen collection.

FIGS. 15A and B is a first embodiment of a specimen collection devicefor a swab where the specimen collection device has no analyticalcapacity.

DETAILED DESCRIPTION Definitions:

The following definitions are provided as an aid in interpreting theclaims and specification herein. Where works are cited or incorporatedby reference, and any definition contained therein is inconsistent withthat supplied here, the definition used therein shall not supersede orlimit the definition supplied herein.

Fomite: An inanimate object or substance, such as a doorknob, utensil,soap bar, or specimen container, that is capable of transmittinginfectious organisms (broadly bacterial and viral) from one individualto another, typically by hand-to-hand or hand-to-mouth exposure to abiological residue on the surface of the inanimate object or substance.

Test samples: Representative biosamples taken by swab include, forexample: gingival, buccal, and mucosal epithelial materials, saliva,wound exudates, pus, surgical specimens, lung and other respiratorysecretions, nasal secretions, sinus drainage, sputum, blood, urine,medial and inner ear contents, ocular secretions and mucosa, cystcontents, cerebral spinal fluid, stool, diarrhoeal fluid, tears, mammarysecretions, ovarian contents, ascites fluid, mucous, gastric fluid,gastrointestinal contents, urethral discharge, vaginal discharge,vaginal mucosa, synovial fluid, peritoneal fluid, meconium, amnioticfluid, semen, penile discharge, or the like may be presented for testingon a swab. Assay from swabs representative of mucosal secretions andepithelia are acceptable, for example mucosal swabs of the throat,tonsils, gingival, nasal passages, vagina, urethra, rectum, lower colon,and eyes. Besides physiological fluids, samples of water, industrialdischarges, food products, milk, air filtrates, and so forth are alsolikely test specimens. Particularly preferred as samples are biosamplescollected on swabs or tampons, where a tampon is essentially ahandleless swab that is sometimes worn internally before testing.

Biohazard: A biohazard is a material, either biologically active orinanimate, that poses a risk or threat to health. Also included in thiscategory as biohazards, sensu lato, as defined here, are materials oflikely biological origin that are visually, tangibly, or odorouslyobjectionable or repulsive, and those materials which are not in fact athreat, but which potentially are a threat until tested negative.Biohazards include potentially infectious material of any kind, and maycontain infectious agents from multiple biological categories, includingbut limited to, bacteria and viruses, either singly or in one or morecombinations thereof, and microbial products such as toxins.

Bioassay Target Molecule: or “analyte of interest”, or “targetmolecule”, may include a nucleic acid, a protein, an antigen, anantibody, a carbohydrate, a cell component, a lipid, a receptor ligand,a small molecule such as a drug, and so forth. Target nucleic acidsinclude genes, portions of genes, regulatory sequences of genes, mRNAs,rRNAs, tRNAs, siRNAs, cDNA and may be single stranded, double strandedor triple stranded. Some nucleic acid targets have polymorphisms,deletions and alternate splice sequences. Multiple target domains mayexist in a single molecule, for example an immunogen may includemultiple antigenic determinants. An antibody includes variable regions,constant regions, and the Fc region, which is of value in immobilizingantibodies. The microfluidic analytical devices of the present inventionare configured to detect a bioassay target molecule of these sorts,singly or in combinations.

Such bioassay target molecules may be associated with a pathogeniccondition: which is taken as a condition of a mammalian hostcharacterized by the absence of health, i.e., a disease, infirmity,morbidity, or a genetic trait associated with potential morbidity.

Microfluidic cartridge: a “device”, “card”, or “chip” with fluidicstructures and internal channels having microfluidic dimensions. Thesefluidic structures may include chambers, valves, vents, vias, pumps,inlets, nipples, and detection means, for example. Generally,microfluidic channels are fluid passages having at least one internalcross-sectional dimension that is less than about 500 μm and typicallybetween about 0.1 μm and about 500 μm, but we extend the upper limit ofthe range to 600 um because the macroscopic character of the beadsuspensions sometimes used as analytical aids require it. Therefore, asdefined herein, microfluidic channels are fluid passages having at leastone internal cross-sectional dimension that is less than 600 um.

Microfluidic cartridges may be fabricated from various materials usingtechniques such as laser stenciling, embossing, stamping, injectionmolding, masking, etching, and three-dimensional soft lithography.Laminated microfluidic cartridges are further fabricated with adhesiveinterlayers or by adhesiveless bonding techniques, such by thermal orpressure treatment of oriented polypropylene or by ultrasonic welding.The microarchitecture of laminated and molded microfluidic cartridgescan differ according to the limitations of their fabrication methods.

Microfluidic pumps: include for example, bulbs, bellows, diaphragms, orbubbles intended to force movement of fluids, where the substructures ofthe pump have a thicknesses or other dimension of less than 1millimeter. Such pumps include the mechanically actuated recirculatingpumps described in U.S. Pat. No. 6,743,399 to Weigl and US 2005/0106066to Saltsman, commonly assigned to the applicant. Such pumps may berobotically operated or operated by hand. Electroosmotic pumps are alsoprovided. Such pumps can be used in place of external drives to propulsethe flow of solubilized reagents and sample in microfluidic device-basedassays.

Blister pack: an on-board reagent pack or sachet under a deformable (orelastic) diaphragm. Used to deliver reagents by pressurizing thediaphragm and may appose a “sharp”, such as a metal chevron, so thatpressure on the diaphragm ruptures the “pillow” (see pillow). These maybe used with check valves or closable vents to produce directional fluidflow and reagent delivery. Elastic diaphragms are readily obtained frompolyurethane, polysilicone and polybutadiene, and nitrile for example(see elastomer). Deformable, inelastic diaphragms are made withpolyethylene terephthalate (PET), mylar, polypropylene, polycarbonate,or nylon, for example. Other suitable materials for the deformable filminclude parafilm, latex, foil, and polyethylene terephthalate Keyfactors in selecting a deformable film include the yield point and thedeformation relaxation coefficient (elastic modulus).

Use of these devices permits delivery or acceptance of a fluid whileisolating the contents of the microfluidic device from the externalenvironment, and protecting the user from exposure to the fluidcontents.

Single entry: refers to a microfluidic device, card or cartridge thatrequires, or permits, only a single introduction of sample, and theassay is then conducted within a self-contained, sealed system. Suchdevices optionally contain a device for sealing or locking the sampleport and an on-board means for disinfecting the contents of the deviceand any waste following completion of the assay. In one embodiment, thedevice can be discarded after use without special precautions.

Waste chamber or “pack”: is a cavity or chamber that serves as areceptacle for sequestering discharged sample, rinse solution, and wastereagents. Typically also includes a wicking material (see wick). Wastepacks may also be sealed under an elastic isolation membrane sealinglyattached to the body of the microfluidic device. This inner membraneexpands as the bibulous material expands, thus enclosing the wastematerial. The cavity outside the isolation membrane is vented toatmosphere so that the waste material is contained and isolated. Wastepacks may optionally contain dried or liquid sterilants.

Vent: a pore intercommunicating between an internal cavity and theatmosphere. A “sanitary” or “isolation vent” also contains a filterelement that is permeable to gas, but is hydrophobic and resistswetting. Optionally these filter elements have pore diameters of 0.45microns or less. These filters function both in forward and reverseisolation. Filter elements of this type and construction may also beplaced internally, for example to isolate a valve or bellows pump fromthe pneumatic manifold controlling it.

Herein, where a “means for a function” is described, it should beunderstood that the scope of the invention is not limited to the mode ormodes illustrated in the drawings alone, but also encompasses all meansfor performing the function that are described in this specification,and all other means commonly known in the art at the time of filing. A“prior art means” encompasses all means for performing the function asare known to one skilled in the art at the time of filing, including thecumulative knowledge in the art cited herein by reference to a fewexamples.

Means for extracting: refers to various cited elements of a device, suchas a solid substrate, filter, filter plug, bead bed, frit, or column,for capturing target nucleic acids from a biological sample, andincludes the cumulative knowledge in the art cited herein. Extractingfurther comprises methods of solubilizing, and relates to theresuspension of cells and tissue from the tip of a swab. This includesmethods, for example, for dissolution of mucous and protein as describedin United States Patent Application 2004/0175695 to Debad. Generally,extraction means include a mechanical pumping component that promotesphysical resuspension by turbulent or near turbulent flow. Such flow maybe reciprocating flow, and may be pulsatile at varying frequencies toachieve the desired resuspension in a reasonable interval of time.Extraction means also include use of detergent-based buffers,sulfhydryl-reducing agents, proteolytics, chaotropes, passivators, andother solubilizing means.

A means for polymerizing, for example, may refer to various species ofmolecular machinery described as polymerases and their cofactors andsubstrates, for example reverse transcriptases and TAQ polymerase, andincludes the cumulative knowledge of enzymology cited herein byreference to a few examples.

Means for Amplifying: The grandfather of this art is the “polymerasechain reaction” (referred to as PCR) which is described in detail inU.S. Pat. Nos. 4,683,195, 4,683,202 and 4,800,159, Ausubel et al.Current Protocols in Molecular Biology, John Wiley and Sons, Baltimore,Md. (1989), and in Innis et al., (“PCR Protocols”, Academic Press, Inc.,San Diego Calif., 1990). Polymerase chain reaction methodologies requirethermocycling and are well known in the art. Briefly, in PCR, two primersequences are prepared that are complementary to regions on oppositecomplementary strands of a target sequence. An excess of deoxynucleosidetriphosphates are added to a reaction mixture along with a DNApolymerase, e.g., Taq polymerase. If the target sequence is present in asample, the primers will bind to the target and the polymerase willcause the primers to be extended along the marker sequence by adding onnucleotides. By raising and lowering the temperature of the reactionmixture, the extended primers will dissociate from the template to formreaction products, excess primers will bind to the template and to thereaction products and the process is repeated. By adding fluorescentintercalating agents, PCR products can be detected in real time.

Other amplification protocols include LAMP (loop-mediated isothermalamplification of DNA) reverse transcription polymerase chain reaction(RT-PCR), ligase chain reaction (“LCR”), transcription-basedamplification systems (TAS), including nucleic acid sequence basedamplification (NASBA), “Rolling Circle”, “RACE” and “one-sided PCR”.

These various non-PCR amplification protocols have various advantages indiagnostic assays, but PCR remains the workhorse in the molecularbiology laboratory and in clinical diagnostics. Embodiments disclosedhere for microfluidic PCR should be considered representative andexemplary of a general class of microfluidic devices capable ofexecuting one or various amplification protocols.

Means for detecting: as used herein, refers to an apparatus fordisplaying an endpoint, i.e., the result of an assay, and may include adetection channel and test pads, and a means for evaluation of adetection endpoint. Detection endpoints are evaluated by an observervisually in a test field, or by a machine equipped with aspectrophotometer, fluorometer, luminometer, photomultiplier tube,photodiode, nephlometer, photon counter, voltmeter, ammeter, pH meter,capacitative sensor, radio-frequency transmitter, magnetoresistometer,or Hall-effect device. Magnetic particles, beads and microspheres haingor impregnated color or having a higher diffraction index may be used tofacilitate visual or machine-enhanced detection of an assay endpoint.Magnifying lenses in the cover plate, optical filters, colored fluidsand labeling may be used to improve detection and interpretation ofassay results. Means for detection of magnetic particles, beads andmicrospheres may also include embedded or coated “labels” or “tags” suchas, but not limited to, dyes such as chromophores and fluorophores;radio frequency tags, plasmon resonance, spintronic, radiolabel, Ramanscattering, chemoluminescence, or inductive moment as are known in theprior art. Colloidal particles with unique chromogenic signaturesdepending on their self-association are also anticipated to providedetectable endpoints. QDots, such as CdSe coated with ZnS, decorated onmagnetic beads, or amalgamations of QDots and paramagnetic Fe304microparticles, optionally in a sol gel microparticulate matrix orprepared in a reverse emulsion, are a convenient method of improving thesensitivity of an assay of the present invention, thereby permittingsmaller test pads and larger arrays. Fluorescence quenching detectionendpoints are also anticipated. A variety of substrate and productchromophores associated with enzyme-linked immunoassays are also wellknown in the art and provide a means for amplifying a detection signalso as to improve the sensitivity of the assay, for example“up-converting” fluorophores. Detection systems are optionallyqualitative, quantitative or semi-quantitative. Visual detection ispreferred for its simplicity, however detection means can involve visualdetection, machine detection, manual detection or automated detection.

Means for isolation include impermeable cartridge body, gas permeablehydrophobic venting, bibulous padding in waste chamber, disinfectant inwaste chamber, elastomeric membrane separating pneumatic actuator fromblister pack, valve with elastomeric membrane actuated by suctionpressure, suction pressure in said sample entry port, on-board reagentpack, self-locking single-entry sample port, gasketed closure, anddisposable external skin or skins Isolation refers both to theprotection of the user from potentially biohazardous specimens, and tothe protection of the specimen from contamination by the user or byforeign environmental materials. Closure means, or “means for sealinglyclosing”, include caps, lids, threaded closures, “ziplock” closures,ball valves, gasketed closures, gaskets, seals, snap caps of all sorts,bungs, corks, stoppers, lip seals, press seals, adhesive seals,waterproof seals, single-entry seals, tamper-proof seals, locking seals,track-slidable sealable covers, compression seals, one-way valves,spring-loaded valves, spring-loaded lids, septa, tee-valves,snap-locking closures in general, piston-valves, double-reed valves,diaphragm closures, hinged closures, folding closures, Luer lockclosures, and so forth.

Unless the context requires otherwise, throughout the specification andclaims which follow, the word “comprise” and variations thereof, suchas, “comprises” and “comprising” are to be construed in an open,inclusive sense, that is, as “including, but not limited to”.

“Conventional” is a term designating that which is known in the priorart to which this invention relates.

“About” and “generally” are broadening expressions of inexactitude,describing a condition of being “more or less”, “approximately”, or“almost” in the sense of “just about”, where variation would beinsignificant, obvious, or of equivalent utility or function, andfurther indicating the existence of obvious minor exceptions to a norm,rule or limit.

Reference throughout this specification to “one embodiment” or “anembodiment” means that a particular feature, structure or characteristicdescribed in connection with the embodiment is included in at least oneembodiment of the present invention. Thus, the appearances of thephrases “in one embodiment” or “in an embodiment” in various placesthroughout this specification are not necessarily all referring to thesame embodiment. Furthermore, the particular features, structures, orcharacteristics may be combined in any suitable manner in one or moreembodiments. Turning now to the figures, FIG. 1 is a conceptual view ofa microfluidic analytical device (1) with integrated sanitary swabcollection features. The device, which is hand sized, is provided withupper and lower disposable external skins (2, lower not shown). Tabs(4,5) assist in peeling off the skins. These skins are removed after thespecimen collection process is completed. Also shown is the swabreceiving orifice (6) and sliding closure (7) in the open position forreceiving a swab. The closure is provided with a seal and track guide(8) whereby the closure is slid into position sealingly covering theswab receiving orifice. The closure is textured with ribs (9) to aid thethumb in moving from left to right (as shown here) in order toeffectuate swab capture within the device. The card body (10) is boundedby external surfaces (11).

FIG. 2 is a representation of a swab (20) as would be used in anembodiment of the invention. The swab comprises a shaft (21) with handleportion (22), neck portion (23), frangible breakaway notch (24), and tip(25) mounted at the distal end of the shaft. The shaft may be of variousshapes or materials. Shaft materials include polypropylene,polyurethane, polycarbonate, polyethylene terephthalate, and otherpolyesters. Also conceived are polyimides such as nylon and naturalfibers such as pine, bamboo, compressed paper, and so forth.

The tip may be of various shapes or materials. Preferred swab shapesinclude a pipe-cleaner shape of bristles, a spade shape with sponge pad,and a “bud” shape with fiber bat. Non-limiting examples of syntheticfiber materials useful in forming swabs include acetate fibers, aramidefibers, polyamide fibers, e.g. nylons, polyester fibers, e.g.polyethylene terephthalate fibers (PET), polyolefin fibers, e.g.polypropylene and polyethylene fibers, polyvinyl alcohol fibers,polyurethane fibers or foams, and mixtures thereof. Further suitablesynthetic fibers include bi- or tricomponent fibers such as PE/PET- orPP/PE fibers. These fibers can for example be so-called core-sheath-,side-by-side- or island-in-the-sea type fibers, as may be useful inselected applications. Lyocell fibers are also useful. Non-syntheticmaterials include woven paper or cotton. Fiber chemistry is generallychosen to be compatible with extraction or analytical chemistries.

Swab fibers may be interlaid, either knitted or randomly entwined.Interlaid webs or fabrics have been formed from many processes, such as,for example, meltblowing processes, spunbonding processes, and bondedcarded web processes. In particular embodiments, interlaid swabmaterials as utilized in the present invention are produced frompolymers, such as, for example, polyethylene or polypropylene. The swabfibers optionally may be made from interbonded fibers, for example as ofthermoplastic fibers. The term “fibers” as used herein refers to a broadrange of thermoplastic members that can be used to form a nonwovenfabric, including members having defined lengths like staple fibers,meltblown fibers that show a beginning and an end, filaments havingendless or continuous lengths, and the like. For example, and withoutlimiting the generality of the foregoing, thermoplastic polymers such aspolyolefins including polyethylene, polypropylene as well as polystyrenecan be used as may be polyesters including polyethylene terephthalate,and polyamides including nylons. Also useful are other thermoplasticpolymers such as those which are elastomeric including elastomericpolyurethanes and block copolymers. Compatible blends of any of theforegoing may also be used. In addition, additives such as wax, fillers,and the like may be incorporated in amounts consistent with the fiberforming process used to achieve desired results. Other fiber or filamentforming materials will suggest themselves to those skilled in the art.Bicomponent fibers may be also used. The fibers may also be formed fromsolution, and examples include viscose. It is only essential that thecomposition be capable of spinning into filaments or fibers of some formthat can be deposited onto a forming surface and thermally formed orinterbonded in a manner dependent upon the forming surface. The swab tipmay comprise a sponge element.

FIG. 3 shows a representative device for swab capture and analysis. FIG.3A is a plan view of the top surface of the device, showing plane ofsection 3B and the location of the swab receiving orifice and sealingclosure. In FIG. 3A, the device body 10 and exterior surfaces 11 areagain shown.

FIG. 3B is a view of the internal workings of a representative device(30), showing a section through the device solid body interior (31),with captive swab tip (32) in swab receiving chamber (33), also termedherein an “internal hollow volume”. In this view, closure (34) andgasket (35) form a liquid-tight seal over the swab receiving chamber 33.Also shown in schematic form are the elements of an on-board nucleicacid assay. Generally, at least one valve (37) will separate theinternal hollow volume of the device body into at least twocompartments, one for the sample receiving chamber and the other theanalytical microfluidics compartment or circuit (dotted lines witharrows, 42). Other valves (38) may also be used to add functionality tothe microfluidic circuit. Any valve known in the art may be used.On-board microfluidic elements for a nucleic acid assay include at leastone microfluidic channel (39), and optionally provision for reagentpacks such as for lysis reagent and extract reagents (40,41), and anoptional microfluidic nucleic acid assay circuit (42), shownschematically. In this embodiment, the internal hollow volume comprisesa first compartment for receiving the swab (33) and a second compartment(42, dotted lines) for performing a fluidic operation on the sample,such as a sample preparation step or a sample analysis such as PCR.Generally, the first and second compartments are joined by a valved (37)microfluidic channel (39). This channel provides for fluidic connectionbetween the compartments so that reagent and sample may beinterechanged. Other compartments such as waste compartment (36) mayalso be provided. Variants of the illustrated microfluidic circuit forjoining the compartments and exchanging fluids between the compartmentsare readily within the scope of the invention. Sample processing stepscould include extraction of the biological material and lysis of cellsof interest, followed by filtration and entry of the filtrate into anucleic acid capture and elution module. Steps of capture, elution,amplification and detection are indicated without detail. Mesoscaledevices for amplification and detection of a nucleic acid in a samplewere first described in 1992 (U.S. Pat. No. 5,498,392 to Wilding,“Mesoscale Polynucleotide Amplification Device and Method”) andconventional mechanisms are known to those skilled in the art. Thesedevices include various filters, pumps, vents, microfluidic channels,valves, and so forth. The device also optionally includes a displaycapability, although this function could be a simple visual indicator,or could be a complex interaction between the device and a docking siteon an instrument that examines fluorescence of an array or a lateralflow strip, and so forth. Therefore, both stand-alone manual diagnosticapplications and automated or semi-automated applications are envisaged.The inner workings of these devices are defined in various embodimentsof the prior art. It should be noted that the claimed invention is notlimited to a particular embodiment of the inner workings, and thatapplications for devices used in performing chemical or immunoassays arealso anticipated. Devices may be built to assay for bioassay targetmolecules indicative of pathological conditions and biological threatsof any kind

Sealing closure 34 comprises a gasket or gasket layer 35. In thisembodiment, the guide track 8 serves also to force a tight seal betweenthe gasket material and the swab receiving orifice 6, thus forming afluid-tight seal over swab capture chamber 33. Following capture, theswab is treated by flowing extraction reagent or buffer in and out ofthe swab receiving chamber. The extraction buffer may includedetergents, solvents such as water, and water in combination with DMSO,NMP, DMF, Formamide, THF, and detergents, co-detergents, cosolvents,proteolytics, sulfhydryl-reducing agents such as n-acetyl-cysteine anddithiothreitol, selective nucleases, mucopolysaccharidases, cellulases,proteases, and the like. A discussion of mucolytics is provided inUnited States Patent Application 2004/0175695 to Debad. Mechanicalagitation is important, and may be enhanced by sonication, such as withpiezoelectric transducers. For reciprocal flow, air in the chamber canbe vented through the waste sequestration chamber or at a secondary ventsite. Optionally, the swab receiving chamber may contain active pumpelements in tandem pairs, operating in alternation by positive andnegative displacement, so that venting is not required. The structure ofthese paired pump elements consists of elastomeric or flexiblediaphragms and the operation requires merely that as the diaphragm ofone pump element is compressed, the other diaphragm is distended, sothat the fluid is forced back and forth between the two pump elements.The diaphragms may be operated manually, hydraulically,electrostatically, magnetically, or pneumatically as is known in theart.

An important capacity of any such device is the sequestration of medicalwaste. The device will typically contain buffer and bioactive reagentsfor sample processing and analysis and all such material is best viewedas biohazardous. Ideally, all such waste is retained in the sealed bodyof the device and can be disposed of without hazard by autoclaving orincinerating the device itself. Shown here is a waste chamber (36) thatwould in operation be vented. Such vents as are permeable to air but notto liquid are well known. Added isolation is possible using a flexiblediaphragm as described in co-assigned US Patent Document “IntegratedNucleic Acid Assays”, where fully operative details of assay systems ofthis sort are disclosed, and which is herein incorporated in full byreference. Also useful are absorbent bats.

Preferably, the devices are self-contained and contain at least on-boardreagent for conducting the analysis. In some cases the reagent is afluid, for example an extraction buffer or a lysis reagent, but in othercases the reagent is a dried biological, for example a primer mix, anantibody, a polymerase, a divalent cation, or a dried weak acid and itssalt. By designing the device to be self-contained, single entry use atthe point-of-care is enabled. Liquid reagent storage may be achieved bysupplying the reagents in sachets, which are ruptured when needed, bymethods known in the art. These methods typically supply a sharp uponwhich the sachet is compressed so that it ruptures. Compression of thesachet may be by manual means or by pneumatic means.

FIG. 4 shows an exploded view of disposable external skins (2,3) appliedto a device body (45). Here, both the upper skin (2) and lower skin (3)are shown. A ribbed surface (44) is provided for gripping the device.These skins may be applied as decals. The upper and lower skins may bemade from a flexible plastic film or sheet, such as polyethylene, vinyl,polyvinyl chloride, PET or polyurethane, and are typically applied tothe device with a removable, pressure sensitive adhesive that can beremoved without residue. Candidate commercially available films include3M™ Scotchcal™ Graphic Film Series 3470 or 3M™ Scotchcal™ Graphic FilmSeries 8000 available from 3M (St. Paul. Minn.) and adhesives includeROBOND™ PS-8211 latexes available from Rohm And Haas (Philadephia, Pa.).Other suitable decal materials include paper sheet, waxed paper sheet,and fiber/plastic or plastic/plastic composite sheets or films, such aspolyethylene film bonded over cloth scrim. These sheets or films aretypically printed with graphics and written instructions for the user.Optionally the instructions are printed onto the device body and thefilm cover is transparent. The adhesive is typically an acrylatederivative. Examples of repositionable and removable adhesives areemulsified polymers made from “soft” monomers such as n-butyl acrylate,isooctyl acrylate, or the like, or ionomeric copolymers made from a softcomponent, such as isobutylene, n-butyl acrylate, isooctyl acrylate,ethyl hexyl acrylate, or the like; in combination with a polar monomersuch as acrylic acid, acrylonitrile, acrylamide, methacrylic acid,methyl methacrylate, trimethylamine methacrylimide, trimethylaminep-vinyl benzimide, ammonium acrylate, sodium acrylate,N,N-dimethyl-N-(.beta.-methacryloxyethyl)ammonium propionate betaine,1,1-dimethyl-1-(2-hydroxypropyl)amine methacrylimide,4,4,9-trimethyl-4-azonia-7-oxo-8-oxa-9-decene-1-sulphonate,1,1-dimethyl-1-(2,3-dihydroxypropyl)amine methacrylimide, and maleicanhydride or the like. Non-spherical polyacrylate adhesives arecommercially available, for example, as the Rohm and Haas Rhoplex™ lineof adhesives. The adhesive applied to the film is typicallyrepositionable or removable without residue, the adhesive may beselected from any adhesive that may be repeatably adhered to and removedfrom a substrate without substantial loss of adhesion capability. Anexample of such an adhesive is disclosed in U.S. Pat. No. 3,691,140 toSilver, which relates to solid tacky microspheres. Preferred adhesivesare water resistant when dry. Repositionable adhesives are also known inwhich microspheres contained in the adhesive are non-tacky. A disclosureof this type of adhesive is provided in U.S. Pat. No. 4,735,837 toMiyasaka, which describes removable adhesives containing elasticmicro-balls with the desired properties. The decal to be applied to thedevice is typically supplied on a release liner and has good moistureand chemical resistance and the adhesive has a working life of greaterthan 6 months. The decal may be a composite multilayered sheet toachieve these objectives. Multilayered decals variously fabricated fromoverlayer, liquid crystalline polymer, plastic, silicone, rubber,thermoplastic, paper, interlaid fiber, underlayer, microporous plastic,backing, scrim, cloth, and adhesive are anticipated for this use.

FIG. 5 shows a representation of how a disposable protective cover canbe applied using tubestock of heatshrink plastic (50), as is readilycommercially available. Once the device is inside a suitable length ofthe heatshrink material, heat is applied to form the coverlayer to theshape of the device. The swab receiving orifice can be provided with anadhesive-backed decal or appliqué that would be removed immediatelybefore use, exposing the orifice, and also serves as a tamper-evidentseal. A tearstrip may similarly be applied to the heatshrink wrapping sothat the entire skin can be removed with a single motion. Candidate heatshrinkable thermoplastic films include those polyethylene compositesdescribed in U.S. Pat. No. 7,235,607, the polyethylene terephthalateesters of U.S. Pat. No. 6,623,821, and the thermoplastics of U.S. Pat.No. 3,655,503, for example.

FIG. 6 describes a similar protective cover, but made out of a softplastic bag such as a polyethylene or polyolefin, or out of paper. Thepaper may be impregnated with a water repellent material or may beabsorbent. The plastic or paper bag (60) is formed to include a malesealing rib (61) that mates with a corresponding female locking groove(62) on the exterior circumference of the device body. A tearstrip isprovided for ease of removal. The swab receiving orifice 6 can beconfigured to a variety of swab dimensions and shapes. When the swab issafely captured within the device, closure 7 is pushed across theopening to seal the device.

The theme is repeated in the composite device (70) of FIG. 7. Here thedisposable outer skin consists of a Styrofoam block or similar expandedmaterial formed by molding, which is fabricated to fit the lower half ofthe device (71), and a partial lid fitted to the upper half of thedevice (72), leaving the swab receiving orifice 6 exposed. A tearstrip(73) serves the dual function of adhering the two halves of the outerskin together during sample collection, and is then torn or peeled awayso that the halves can be separated and the device removed for furtherprocessing or analysis. The tearstrip typically includes a freehangingtab to facilitate this. The lower block and upper lid are discardedafter the device is removed.

Note that the shape of the blocks forming the outer skin 70 is variable.A clamshell formed of right and left halves is equally suitable, as aremore complex interdigitated two part blocks. A single block is useful.The dual block system has the advantage that squeezing pressure appliedto the lower block has the effect of holding the device in place whilethe tear strip and upper lid are removed. The device can then be pulledout of the lower block with clean hands and presents an uncontaminatedexterior, the closure having been pulled over the swab receiving orificefrom its protected position under the upper lid.

FIG. 8 shows a conceptual view of a more general form of the compositesample collection device (80) with disposable outer skin (81). Here thedisposable outer layer material can be a quilted material, a compositeof waterproof and absorbent layers, a diaper, a foil composite, and soforth. The material is knit or fused around the edges into a pouchholding the device, and is torn away at a frangible or pre-weakened tearpoint after the sample is collected. U.S. Pat. No. 4,279,344 describes apackaging laminate which is heat sealable and peelable suitable for thisconstruction.

FIG. 9 is a pictorial representation of the essential features of theswab capture method, and shows a multistep process with steps A-E and arepresentative device (1) and swab (20). In FIG. 9A and B, the swab (20)is oriented to the swab receiving orifice (6) of the device body (1) andthe tip of the swab (25) is inserted into the device. In step C, thehandle (22) is broken away and discarded. The locking closure (7) isthen slid over the orifice (6) to irreversibly capture and seal the swabtip in the device, as shown in FIG. 9D. In step E, the disposableexternal skins, or “decals”, are then peeled away (shown is the upperskin 2 peeling away), refreshing the external surfaces and removing anyextraneous material inadvertently deposited when collecting the sample.The fresh external surfaces are used to label the specimen contents andpatient identification, or optionally a label with that information canbe applied to those surfaces.

FIG. 10 is a block diagram of these steps of the general method for swabcapture. The steps are: collect a specimen on a swab; insert the swabtip into the collection device as designed for receiving the swab, andbreak off the swab handle; seal the swab in the device using a lockingclosure; remove the disposable skin or skins from the external surfacesof the collection device, taking care to avoid contaminating the freshlyexposed surfaces. Optionally, an analysis may then be performed on theswab in the device without further exposure to the biohazardous sample.

Note that the order of the steps is not strictly followed if the swabhandle is broken off and the device sealed after the external skins areremoved, and it may be that handling the device in this way is moreconvenient. However, the preferred method is to capture the swab andseal the device before removing and discarding the external protectiveskins. As claimed, the invention is not limited by the order of thesesteps.

FIG. 11 is a block diagram of a more general method for specimencapture. The specimen is first collected and inserted in a suitablecontainer, the container having been supplied with disposable externalskin or skins; the container is then sealed; and the external skins orskins are removed and discarded.

FIGS. 12A and 12B are overview and detail, respectively, of the tabmembers (4,5) used as a peelaway strip for removing the external skinsof a representative device. As shown in detailed view 12B, the tabs arefreestanding at the edges of the body of the device, and are easilygrasped between finger and thumb. The entire protective film or pad isthen readily peeled away.

FIG. 13 is an alternate embodiment of a combination specimen collectioncontainer and sheath (130), showing an alternate form of the externalprotective skin and internal specimen collection device. Here theanalytical device (131) shown is fitted with internal analytical worksand a user interactive panel and display window.

In use, the body of the sample collection device 131 is encased in anouter sleeve member (132) and cap member (133). The outer sleeve memberis supplied with an endwise swab receiving orifice (134) and internalswab receiving chamber for collecting the swab. A ball valve typeclosure is used to capture and seal the swab in the device and a knob isprovided (135) for rotating the ball valve from open to closed. Thecontrol head 136 may also be rotated, and serves to power aspring-driven pressure source for the pumps, and to initiate the assayprotocol. Assay status is shown in the leftmost window 137. Assayresults are shown in the rightmost window 138.

After the sample is collected, the outside protective sleeve 132 isremoved and the sample receiving chamber is closed with the ball valve135. The cap can then be removed and the apparatus is generally free ofexternal contamination. The sample entry end can be covered. The controlhead is then rotated and the assay commenced. In a few minutes, theassay result is read in the display window. Status and validity of theassay is displayed in the left panel. Optionally, the device can beinserted into a machine and the assay conducted by machine-aided powerand control. The outer sleeve and cap are discarded as contaminatedmedical waste. At the completion of the assay, the device is alsodiscarded along with its entrained specimen.

Note that the embodiment is illustrative of a general concept, and isnot limited by its specificity. The outside protective sleeves aredisposable external skins The sleeves may be replaced by decals asdescribed in FIG. 4, wherein the decals are adapted for a cylindricalbody form. Similarly, the disposable protective overlayer may be asprovided in FIGS. 5-8.

This device is also suitable for collection of tampons, which lack thehandle characteristic of swabs. The tampon, however, must be insertedinto the swab receiving orifice with tweezers or by other means and theorifice must be dimensioned appropriately. Tampons are useful samplecollection devices, and their use is hereby taken within the scope ofthe invention described herein.

This device is conceived as part of a kit, the kit consisting of asterile swab, the combination specimen collection device and sheath 130,and a tray. The tray optionally may also contain surgical gloves,instructions, and labeling aids.

A variation of composite device 130 is shown in FIG. 14. Here the sampleis inserted through orifice (141) in external disposable cap (142) intodevice (140), the body of which contains a sample receiving chamber withthreaded neck (143). After the sample is deposited in the device, thecap 142 is immediately removed and a clean, sterile lid (not shown) isthreaded onto the neck. The device body thus functions as bottle.Holding the assembly by the clean lid, the lower outside protectivesheath (144) is then removed.

The external surfaces of the device are now clean and safely handledwithout gloves. Objectionable materials deposited on the outside sheathare discarded along with the disposable sheaths, which function as anexternal protective skin.

In this embodiment, the operator then presses the start button (146);the instrument cycles, its status continuously displayed in status bar147, and the raw data is read from nucleic acid hybridization array 148.The machine is placed under a modified bar code reader or strip readerand the data is electronically displayed on the reader and transmittedas an electronic medical record to the patient's chart.

These various analytical features are not presently viewed aslimitations of the present invention. The present invention relates tomethods and devices for collecting specimens and for analyzing specimensin which a pre-formed disposable external skin is removed from thecollection device or sample holder after the specimen is deposited init.

Thus in FIG. 15, a swab collection container is shown with no analyticalcapabilities. The composite swab holder consists of an internal bottleand an external skin or sheath, so that after the swab is collected andsealed within the internal bottle, the external sheath is removed andthe swab in its bottle, or other sealed vessel, is safely transportedand handled with the assurance that any biohazardous external residueshave been disposed of with the external sheath.

Swab collection container (150) is shown in FIG. 15A. Internal swabcollection container (151) is shown in FIG. 15B. The two figuresillustrate essentially a “before” and “after”, wherein the device issupplied as shown in FIG. 15A without collected swab, and in FIG. 15Bwith collected swab. The steps involve capturing the swab and removal ofthe external skins, so that the product of the method is the slender,clean swab holder shown in FIG. 15B.

As supplied, the swab collection container 150 has a swab receiving port(153) formed of disposable funnel (154) and barbed lip (164) of theinternal swab receiving channel (156), also termed herein an “internalhollow volume (156)”. The temporary shipping cap (160) is first removedand the swab is inserted tip-down into the internal hollow volume (156).Note that the disposable funnel serves to protect the barbed rim (164)of the internal sheathed tube (157) from contamination with specimenresidues. Following collection of the swab and placement within theinner tube, the sealing strip, or tear strip (168), is removed and theupper protective skin (155) is lifted up and away from the device, alongwith the disposable funnel 154, both of which are discarded. Thisexposes the uppermost bezeled rim 164 of the inner cylinder. Now, asshown in FIG. 15B, a sealing closure (165) with locking lip or flange(166) and plug (167) can be locked in place over the barbed bezel of theinner cylinder, and the outer lower protective sheath 159 is slid offthe inner cylinder and discarded. The sealing closure is suppliedseparately. After these steps, the swab is now isolated within theinternal hollow volume 156, separated from the external surfaces (169)by closure 165, and the external surfaces are as clean as supplied bythe factory.

Note that the removal of the outer shells is a two part process. With agloved hand, the contaminated outer shell is grasped and the upper shellis removed. A clean hand is then used to install the closure, and theupper part of the inner cylinder is held while the lower shell isremoved. The final specimen container is now free of contamination andcan be handled without gloves. To later gain access to the swab,fracture lines such as described in U.S. Pat. No. 6,516,947 may beformed in the internal cylinder, which can be formed generally asdescribed in FIG. 1 of that publication. In that way, it is nevernecessary to touch the directly swab again. Alternatively, the closureof the device of FIG. 15 can be a threaded closure, and the internalcylinder may be formed with a mating threaded rim and sealing flange.Various combinations are anticipated.

If a patient were to collect the sample, we envisage that the patientwill place the swab in the device and return it, outer shell intact, toa healthcare professional or laboratory technician. The technician willthen complete the steps of removing the upper shell, inserting andsealing the cap, and then removing the lower shell, taking care to avoidcontaminating the external surfaces of the inner cylinder during theprocess. Between steps of the process, the device 150 may be stood onits base, which can be formed with a foot as would be useful forstability.

A kit for this process may contain, in a tray, the device 150, a swab20, and a closure, along with any instructions and labeling.

It will be appreciated by persons skilled in the art that numerousvariations, combinations of elements, and/or modifications may be madeto the invention as shown in the specific embodiments without departingfrom the spirit or scope of the invention as broadly described. Thepresent embodiments are, therefore, to be considered in all respects asillustrative and not restrictive.

EXAMPLES Example 1

A swab is provided in a sterile packet, the shaft of the swab beingformed with a notch separating the handle from the sampling tip. Theswab is rubbed in the gingiva separating the teeth from the gums of achild and inserted into a collection device of the invention. The swabhandle is bent vigorously so that it breaks at the notch, releasing theswab tip with specimen into the device. The swab insertion channel isthen covered with a sliding closure that rides in tracks in the housing,and sealed irreversibly, the sliding closure having a ratchetedunderside which mates and locks over a locking tooth or spur on the bodyof the device. The professional then removes a protective external skinfrom the device, taking care not to contaminate the freshly exposedsurfaces, and hands the device to an aide for processing.

Example 2

A swab is provided in a sterile envelope, the shaft of the swab beingformed of a material suitable for cutting with a blade. The patient isasked to provide a self-collected specimen of the vaginal mucosa and isgiven instructions. The patient collects the sample and inserts the softtip of the swab into the sample collection device that was provided. Thepatient hands the device to a health professional, who takes it withgloved hands. The health professional closes the cover of the device,cutting free the swab handle and discarding it, and then removes thedisposable external skins on the device, taking care not to contaminatethe freshly exposed surfaces. After removing the skins, the healthprofessional inserts the device into a semi-automated analyticalapparatus and completes the assay. The result is read and the devicewith sample is then discarded. The analytical apparatus is equipped withnetworking capability so as to transmit identifying and “smart”electronic data as an electronic medical record to a database on aserver.

1. A sample collection device, the device comprising: a) a body havingan external surface and an internal hollow volume; b) a removabledisposable external skin layer or shell covering a first portion of theexternal surface of the body and dividing the external surface into acovered area and an uncovered area, the uncovered area defining a swabreceiving orifice for inserting a swab into the internal hollow volume,the swab receiving orifice compatible with a sealable closure that maybe sealably closed over the swab receiving orifice; and, wherein theremovable disposable external skin layer or shell comprises a firstgripping surface and the uncovered area comprises a second grippingsurface, said first gripping surface having means for removing saidexternal skin layer or shell from said body.
 2. The sample collectiondevice of claim 1, wherein the external skin layer or shell is a decalhaving a pressure sensitive adhesive that can be removed without leavinga residue.
 3. The sample collection device of claim 1, wherein theexternal skin layer or shell comprises a first part and a second part.4. The sample collection device of claim 3, wherein the first part andthe second part are joined by a tear strip.
 5. The sample collectiondevice of claim 1, wherein the external skin layer or shell is a foamsheath shaped with a receptacle covering the first portion of theexternal surface of the body.
 6. The sample collection device of claim1, wherein the external skin layer or shell is a foam block encasing thefirst portion of the external surface of the body.
 7. The samplecollection device of claim 6, wherein the foam block is divided into afirst member and a second member, wherein the first gripping surface isdefined on the first member.
 8. The sample collection device of claim 1,wherein said second gripping surface comprises a cap adapted to hold thebody with a clean hand while removing and discarding the external skinlayer or shell.
 9. The sample collection device of claim 1, furthercomprising a swab, wherein the swab comprises a handle with frangibleneck and an absorbent tip for collecting the sample.
 10. The samplecollection device of claim 1, wherein the swab is a tampon.
 11. Thesample collection device of claim 1, wherein the body comprises internalworkings for processing the sample prior to performing an analysis onthe sample.
 12. The sample collection device of claim 1, wherein thebody comprises internal workings for analyzing the sample.